Michaelsen T E, Kolberg J
Acta Pathol Microbiol Immunol Scand C. 1984 Feb;92(1):25-35. doi: 10.1111/j.1699-0463.1984.tb00048.x.
Antibodies were made against the two-chain lectin Lath-O from the seeds of Lathyrus odoratus as well as its isolated light (alpha) and heavy (beta) chains. These antibodies were used to antigenically compare the Lath-O with other two-chain lectins like Lath-S, lentil and Vicia cracca glc specific and with one-chain lectins like Con A, PHA, peanut and soybean. Sensitive ELISA tests showed that there was a distinct antigenic cross-reaction between the one-chain and two-chain lectins both by using antibodies against Lath-O whole molecules and its isolated beta-chains, while the anti-Lath-O alpha-chain antibodies barely reacted at all even with Lath-O alpha-chains. The antibodies against Lath-O whole molecules also strongly inhibited the mitogenic responses induced by two-chain lectins. Antibodies reacting inside the carbohydrate-binding site of the lectins were isolated by eluting them with glucose from lectin-Sepharose columns and antibodies reacting outside the carbohydrate-binding site were subsequently eluted with guanidine or buffer with low pH. The two antibody fractions were separately used to antigenically compare one-chain and two-chain lectins, demonstrating that both kinds of antibodies showed cross-reaction between one-chain and two-chain lectins. Both the antibodies presumably reacting inside the carbohydrate-binding site of the lectins and those reacting outside the carbohydrate-binding site inhibited haemagglutination activity of two-chain lectins, while haemagglutination by one-chain lectins was unaffected. The carbohydrate-binding site specificity of the glucose-eluted antibodies was assessed by glucose-inhibition in ELISA tests. Furthermore, galactose neither inhibited in these tests nor eluted antibodies from lectin-Sepharose columns. Control experiments were also performed with normal rabbit IgG.
制备了针对香豌豆种子中的双链凝集素Lath - O及其分离的轻链（α链）和重链（β链）的抗体。这些抗体用于对Lath - O与其他双链凝集素（如Lath - S、小扁豆和野豌豆glc特异性凝集素）以及单链凝集素（如伴刀豆球蛋白A、植物血凝素、花生凝集素和大豆凝集素）进行抗原性比较。灵敏的酶联免疫吸附测定（ELISA）试验表明，使用抗Lath - O全分子及其分离的β链抗体时，单链和双链凝集素之间存在明显的抗原交叉反应，而抗Lath - O α链抗体即使与Lath - O α链也几乎不发生反应。抗Lath - O全分子抗体也强烈抑制双链凝集素诱导的促有丝分裂反应。通过用葡萄糖从凝集素 - 琼脂糖柱上洗脱，分离出在凝集素碳水化合物结合位点内部起反应的抗体，随后用胍或低pH缓冲液洗脱在碳水化合物结合位点外部起反应的抗体。将这两种抗体组分分别用于对单链和双链凝集素进行抗原性比较，结果表明这两种抗体在单链和双链凝集素之间均显示出交叉反应。推测在凝集素碳水化合物结合位点内部起反应的抗体和在该位点外部起反应的抗体均抑制双链凝集素的血凝活性，而单链凝集素的血凝不受影响。通过ELISA试验中的葡萄糖抑制来评估葡萄糖洗脱抗体的碳水化合物结合位点特异性。此外，半乳糖在这些试验中既不产生抑制作用，也不能从凝集素 - 琼脂糖柱上洗脱抗体。还使用正常兔免疫球蛋白G进行了对照实验。
